Insight Into the Hidden Bacterial Diversity of Lake Balaton, Hungary

Biologia Futura (2020) 71:383–391 https://doi.org/10.1007/s42977-020-00040-6

ORIGINAL PAPER

Insight into the hidden bacterial diversity of Lake Balaton, Hungary

E. Tóth1 · M. Toumi1 · R. Farkas1 · K. Takáts1 · Cs. Somodi1 · É. Ács2,3

Received: 22 May 2020 / Accepted: 17 August 2020 / Published online: 7 September 2020 © The Author(s) 2020

Abstract In the present study, the prokaryotic community structure of the water of Lake Balaton was investigated at the littoral region of three diferent points (Tihany, Balatonmáriafürdő and Keszthely) by cultivation independent methods [next-generation sequencing (NGS), specifc PCRs and microscopy cell counting] to check the hidden microbial diversity of the lake. The taxon-specifc PCRs did not show pathogenic bacteria but at Keszthely and Máriafürdő sites extended spectrum beta- lactamase-producing microorganisms could be detected. The bacterial as well as archaeal diversity of the water was high even when many taxa are still uncultivable. Based on NGS, the bacterial communities were dominated by Proteobacteria, Bacteroidetes and Actinobacteria, while the most frequent Archaea belonged to Woesearchaeia (Nanoarchaeota). The ratio of the detected taxa difered among the samples. Three diferent types of phototrophic groups appeared: Cyanobacteria (oxygenic phototrophic organisms), Chlorofexi (anaerobic, organotrophic bacteria) and the aerobic, anoxic photoheterotrophic group (AAPs). Members of Firmicutes appeared only with low abundance, and Enterobacteriales (order within Proteobac- teria) were present also only in low numbers in all samples.

Keywords Lake Balaton · Bacterial diversity · Archaea · Bacteria · Next-generation sequencing (NGS)

Introduction (Istvánovics et al. 2008; Bolla et al. 2010; Hatvani et al. 2014; Maasz et al. 2019). Lake Balaton is the biggest lake of Central Europe, a shal- The frst microbiological investigations aimed at studying low water ditch, with an average depth of 3.0–3.6 m. The the prokaryotic diversity of the lake showed that members open water of the lake is almost permanently homogenized of the genus Bacillus (Firmicutes) are dominant at some by wind, resulting concomitant resuspension of the sedi- regions (Langó 1982). Diferent Actinobacteria were also ment. The lake has a great importance also due to its tour- isolated from the water of Keszthely Bay, e.g. Streptomy- istical use. The quality of the water is regularly tested, and ces and Micromonospora species (Farkas 1982). In 1985, monitoring has started already in the 1960s (Szabó 1999; the presence of Enterobacteria (indicating faecal pollution) Kiss et al. 2005). In 2006, Hungary joined the water quality was found in Keszthely Bay, and Enterobacter agglomer- assessments in line with the EU Water Framework Direc- ans, Escherichia coli, Kluyvera cryocrescens and Klebsiella tive (EU WFD). In it, the defnitions of biological variables oxytoca were detected in high number among the cultivated were given a much stronger role than before. Moreover, till bacterial strains (Bognár, 1990). that time many studies had revealed the state of the water Tóth (1995) has studied the bacterial communities of the open water in the Balatonfüred region and found the dominance of many Gram-negative organisms belonging to the genera Aeromonas, Pseudomonas and Acinetobacter. Bacte- * E. Tóth rial partners of Eudiaptomus gracilis (important member of [email protected] the zooplankton of the lake) were also studied (Homonnay 1 Department of Microbiology, Eötvös Loránd University, et al. 2011) as well as the microbiology of big-headed carps Budapest, Hungary (Borsodi et al. 2017). The frst detailed reports about aerobic 2 MTA Centre for Ecological Research, Danube Research anoxygenic phototrophs (AAPs) of the lake showed that their Institute, Budapest, Hungary abundance as compared to heterotrophic bacteria is between 3 National University of Public Service, Faculty of Water 1 and 7% (Szabó-Tugyi et al. 2019). Sciences, Baja, Hungary

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All of these studies are very important, but most of them DNA extraction from the water samples were based on cultivation techniques, and it is known that less than 1% of bacteria are cultivable from natural aquatic For DNA extraction, 0.25L water samples were fil- habitats (Amann et al. 1995). tered through 0.22 μm pore size, mixed cellulose fil- The aim of the present study was to reveal the prokaryotic ter (type GSWP; Millipore, Billerica, MA, USA) using community structure of the water of the lake at the littoral the Ultraclean ® PowerSoil DNA Isolation Kit (MoBio, region (where the water is subjected also to strong human Carlsbad, CA, USA) according to the manufacturer’s infuence due to bathing) by cultivation independent meth- instructions. ods [next-generation sequencing (NGS) and microscopic cell counting]. By using specifc PCRs, the presence of some hygienically important bacteria was tested. Our results show Specifc PCRs the hidden prokaryotic diversity of the lake which has never been revealed until now, and provide information about the Taxon-specifc PCRs were applied for the following bacte- potential health risks of bathing. rial groups: Pseudomonas spp. and Pseudomonas aeruginosa according to Lavenir et al. (2007), Legionella spp. according to Cloud et al. (2000), Legionella pneumophila Materials and methods according to Fiume et al. (2005), Acinetobacter baumannii according to Tsai et al. (2018) and Stenotrophomonas malt- Sampling ophilia according to Filho et al. (2004). At the sampling sites there were no people at the time of Sampling was carried out in 30. 08. 2018 at three points sampling (it was late pm), but the sites were subjected to of the lake: Tihany (65.91900 N; 17.88903 E), Kesz- bathing in the previous days, therefore, the bacterial com- thely (46.76014 N; 17.25448 E) and Balatonmáriafürdő munity was tested also for extended spectrum beta-lactamase (46.70725 N; 17.38276 E) (later: Máriafürdő), from the lit- (ESBL)-producing and macrolide-resistant bacteria. toral water region. The water samples (1–1 L) were asep- The presence of ESBL genes was tested by multiplex tically collected into sterilized screw-capped fasks from PCRs in the water samples according to Trung et al. (2015) 10 cm subsurface according to ISO 19458:2006. Samples and macrolide-resistance genes following the protocol were transported to the laboratory in a cooler bag (at 4 °C) described by Zmantar et al. (2011), testing fve genes (ermA, and processed within 4 h after sampling. ermB, ermC, msrA and mefA) simultaneously.

Analysis of physical and chemical parameters Next‑generation DNA sequencing and data analysis

To measure the physical and chemical parameters, the fol- For the identifcation of bacterial and archaeal taxa, the 16S lowing standards were used: MSZ 448-32:1977 (conductiv- rRNA gene of the DNA samples was amplifed with PCRs ity), MSZ ISO 5813:1992 (dissolved oxygen, DOC), MSZ in separate, triplicate reactions using primers with the fol- 1484-13:2009 (nitrate concentration), MSZ 448-13-1983 lowing target-specifc sequences: Bact_341F (5′-CCT ACG (sulphate concentration), MSZ ISO 10260:1993 (chlorophyll GGN GGC WGC AG-3′) and modifed Bact_805R (5′-GAC a). The measures were done by the Central Transdanubian TAC NVG GGT ATC TAA TCC-3′) for bacteria (Herlemann Water Authority, and we got it via the National Directorate et al. 2011), and A519F (5′ -CAG CMG CCG CGG TAA- of Water Management (OVF); data were obtained from the 3′) and Arch855R (5′ -TCC CCC GCC AAT TCC TTT ofce except values of water temperature and pH which were AA-3′) for Archaea (Klindworth et al. 2013). DNA sequenc- measured on site. ing was performed on an Illumina MiSeq platform using MiSeq standard v2 chemistry by the Genomics Core Facil- Determination of microscopy cell counts ity RTSF, Michigan State University, USA. Sequencing and the description of bioinformatic analysis were done accord- Determination of bacterioplankton abundance (micros- ing to Benedek et al. (2019). OTUs were determined at the copy cell counts) was carried out as described by Kéki species-level delineation based on Tindall et al. (2010). et al. (2019) using Nikon80i epifuorescent microscopy Sequence reads were deposited in the NCBI SRA database and NisElements program package. For the investigations, and are accessible through the BioProject PRJNA628507, 10 mL water samples were fltered on sterile polycarbonate Biosample ID SAMN14732963 (Tihany), SAMN14732959 flters (Millipore, Billerica, MA, USA) and fxed with 2% (Máriafürdő) and SAMN14732964 (Keszthely). Diversity paraformaldehyde solution. indices were calculated using mothur (Schloss et al. 2009),

1 3 Biologia Futura (2020) 71:383–391 385 and reads were subsampled to the read number of the sample having the lowest sequence count. Cell count/mL 6.34E+06 6.13E+06 Results 2.75E+06

Physico‑chemical and biological parameters (mg/l 2− 4 94 91 119 The physico-chemical parameters together with the cell SO count values of the samples of the Lake Balaton are given in Table 1. Data of the three sampling sites showed similari- (mg/l) −

ties though some diferences could be detected: the quantity 3 0.1 0.1 0.1 of total organic carbon (TOC), total organic nitrogen (TON), NO total organic phosphorous (TOP) as well as chlorophyll-a ) concentration were the lowest in the Tihany region. Most 3 TOC appeared in dissolved form (DOC), and the water was 5.33 aerated at all sites. 7.7 21.3 Chlorophyll Chlorophyll a (mg/m Specifc PCRs

The taxon-specific PCRs showed (Table 2) that Pseu- 39 66 65 domonas spp. and Legionella spp. were present at all sites, (μg/L) TOP but pathogenic L. pneumophila and P. aeruginosa as well as Acinetobacter baumannii could not be detected. At Kes- zthely and Máriafürdő sites ESBL-producing bacteria could 0.646 0.956 0.766 also be detected. (mg/L) TON

Amplicon sequencing

NGS results showed that diversity indices of Keszthely 7.6 8.5 8.7 and Máriafürdő samples were similar in case of all types DOC (mg/L) of indices, while Tihany sample (despite the fact that cell counts were high even at that site) produced much lower values. The Chao indices (abundance-based estimator of species richness) were 747.5 in case of Tihany, 1433.4 in 7.7 9.6 9 TOC (mg/L) TOC case of Keszthely and 1509 in case of Máriafürdő sample, respectively. The distribution of the diferent bacterial phyla and dominant classes among the samples is given in Fig. 1. 9.14 9.52 10.01 Results show that on the level of phyla the samples are Dissolved (mg/L) oxygen similar: Proteobacteria, Bacteroidetes and Actinobacteria 8.6 8.5 8.4 were the most abundant members of the communities at each pH site and many unclassifed bacteria: Planctomycetes, Cyano- bacteria and Patescibacteria also occurred. Abundance of Firmicutes was relatively low in case of each sample. On 680 610 600 the other hand, the ratio and composition of the diferent Conductivity (μS/cm) lower-level taxa difered among the samples. Most abundant 22 22.3 23.4 orders among Alphaproteobacteria were Rhizobiales, Azos- T °C pirillales, Rhodobacterales, Rhodospirillales and members of SAR11 clade (Azospirillales and Rhodospirillales were present only at Tihany region), among Deltaproteobacteria, in the Balaton summer of 2018 of Lake and cell count values parameters and chemical of the determinationResults of physical Bdellovibrionales, Myxococcales and many sulphate-reduc- 1 Table site Sampling Tihany Máriafürdő ing bacteria (SRB) appeared, while Gammaproteobacteria Keszthely

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Table 2 Results of the taxon-specifc, ESBL-producing and macrolide resistance genes by PCRs Sampling site Pseu- P. aeruginosa Legionella L. pneu- Stenotrophomonas Acinetobacter ESBL-pro- Macrolide domonas spp. mophila maltophilia baumannii duction resistance spp.

Tihany + − + − + − − − Máriafürdő + − + − + − + − Keszthely + − + − + − + −

100%

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30%

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10%

0% Tihany Máriafürdő Keszthely

Acidobacteria Acnobacteria Armamonadetes Bacteria_unclassiﬁed Bacteroidetes Chlamydiae Chloroﬂexi Cyanobacteria Firmicutes Omnitrophicaeota Gemmamonadetes Nitrospirae Patescibacteria Planctomycetes Proteobacteria Verrucomicrobia

Fig. 1 The distribution of the diferent bacterial phyla and the ratio of dominant classes among the samples of Lake Balaton

(giving more than 60% of Proteobacteria at each site) were organotrophic bacteria and the aerobic, anoxic photohet- represented by the abundance of unclassifed Burkholde- erotrophic group (AAPs). Their distribution among the riaceae, Methylococcales (was not present at Tihany sam- samples and the ratio of diferent Cyanobacteria are dis- ple), Betaproteobacteriales, Pseudomonadales and Xan- played in Fig. 2. thomonadales. Enterobacteriales were present only in low Among Cyanobacteria, ratio of Nostocales was high at numbers in all samples. Keszthely and Máriafürdő samples, while members of Syn- From the phylum Actinobacteria members of Micro- echococcales were more abundant in the Tihany region, and trichales, Frankiales and Micrococcales appeared in high they are represented by the dominance of Cyanobium (pico- number, their ratio was higher at Máriafürdő and at Kes- cyanobacterium). Among Chlorofexi, members of the order zthely. Among Bacteroidetes, the orders Chitinophagales, Anaerolineales were the most abundant, but they did not Cytophagales, Flavobacteriales and Sphingobacteriales were appear in Tihany sample. abundant, at Tihany sample, less orders were identifed than In case of Archaea, the diversity indices showed simi- in the other two samples. lar tendencies than in case of bacteria, Chao indices were Three different types of phototrophic metabolisms 108.27 in case of Tihany, 178.9 in case of Keszthely and appeared in the samples: Cyanobacteria are oxygenic 119.02 in case of Máriafürdő sample, respectively, and val- phototrophic organisms, Chloroflexi are anaerobic, ues were much lower than in case of bacteria.

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b 100%

90%

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70% a 100% 60% 90%

80% 50% 70%

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Tihany Máriafürdő Keszthely 0% Chloroflexi Cyanobacteria AAPs Tihany Máriafürdő Keszthely Cyanobacteria_unclassified Vampirovibrionales Leptolyngbyales Nostocales Oxyphotobacteria_unclassified Synechococcales

Fig. 2 a Distribution of phototrophic bacteria among the samples. b Ratio of the diferent Cyanobacteria (Nostocales, Synechococcales, Vam- pirovibrionales, Leptolyngbiales and unclassifed Cyanobacteria) among the samples of Lake Balaton

Fig. 3 Distribution of archaeal 100% phyla among samples of Lake Balaton 90%

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0% Tihany Máriafürdő Keszthely

Alarchaeota Archaea_unclassiﬁed Asgardaeota Crenarchaeota Diapherotrites Euryarchaeota Nanoarchaeaeota Thaumarchaeota

The archaeal phyla appearing at the samples are shown community. Crenarchaeota (e.g. Bathyarchaeia) and Eur- in Fig. 3. yarchaeota (e.g. Methanobacteria and Thermoplasmata) The absolute dominance of Nanoarchaeota was obvi- were very less abundant in Tihany sample, and Asgardeota ous in all samples, and Woesearchaeia dominated each did not even appear there. Unclassifed Archaea were pre- sample among them: representing more than 65% of each sent in all of the three samples.

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Discussion Boeck et al. 2012; Blaak et al. 2014), since these regions are strongly afected by bathers. The quality of the surface waters (e.g. lakes, rivers, If we check the composition of bacterial communities, we streams) is very important, they often serve as basis for can immediately see that the diversity of the lake is much drinking waters or act as touristic sites, also for bathing higher than we have ever thought. NGS results have not only purposes. In the nature, complex systems exist, where shown the presence of many non-cultivable taxa but indi- the characteristics of the given habitat are determined cated also that those bacteria which previously were thought by inside and outside efects, infuenced by interacting to be dominant, occur in the lake only with low frequency. microbial populations as well as anthropogenic efects Since, that our results are based on molecular data com- (e.g. bathing or any contamination). Studies using next- parisons with previous studies is not really applicable, it is generation sequencing (NGS) technologies are providing worth mentioning: members of Firmicutes appeared only new insights into the ecology of microbially mediated pro- with low abundance in the water of the lake at each sampling cesses that infuence freshwater quality. sites (e.g. Bacillus, Exiguobacterium species) though they Among the three sampling sites at Lake Balaton, only were frequently isolated earlier (Langó 1982; Homonnay small diferences could be revealed in the composition of et al. 2011). bacterial communities, and all can probably be explained, Among Proteobacteria, mainly diferent Pseudomonas, e.g. by the small shifts and diferences in the organic mate- Aeromonas and Acinetobacter species were detected pre- rial (TOC, TON, TOP) content or the light access of the viously (Tóth, 1995), their seasonal abundance was also diferent sampling sites. Though the water body is aerobic remarkable. (the dissolved oxygen content was always high), being a The present studies showed that in the water there are shallow lake, the wind can easily stir the whole water col- several bacteria which can be responsible for N 2 fxation umn, so anaerobic microbes can originate also from the (Rhizobiales, Azospirillales, etc.) which phenomenon can sediment. Moreover, microbes can live in microhabitats be very important in low nutrient content environments, where the oxygen concentration can difer strongly that of where the nitrogen is often in limited concentration (How- the water column. arth and Marino 2006). Nitrogen fxation was documented In case of Lake Balaton, based on the chemical and also in case of Lake Balaton earlier (Kovács et al. 2012; biological parameters all the three sampling sites are low Borics et al. 2016), authors revealed that during eutrophi- nutrient content habitat, seemed to be oligotrophic (https:// cation processes nitrogen-fxing, flamentous cyanobacteria www.vizugy.hu/index.php?module=vizstrat&programele became frequent in the western part of the lake. This time mid=149; Borics and Kiss 2015) at time of samplings, also the heterotrophic, free living nitrogen-fxing bacteria though to state it, more investigations should be done (e.g. were identifed. checking macrozoobenthos, macrophyton, etc.). But our Based on literature data, bacteria belonging to SAR11 result shows that even at the end of a summer period, when have an oligotrophic lifestyle with slow but efcient nutrient anthropogenic efect is higher due to bathing, the self- uptake at low substrate concentrations (Salcher et al. 2011). purifcation of the lake is adequate. Studying the seasonal The concentration of TOC was low at each sampling site, dynamics of the phytoplankton, Somogyi et al. (2016) which could have a positive efect on the abundance of sub- revealed a west-east trophic gradient in the longitudinal group III of SAR11. axis in the lake, similar to the previous years. The presence of Bdellovibrionales and Myxococcales In 2018, the tendency is similar, though the cell counts (and even that of Vampirovibrionales) shows that even of bacteria were in the same magnitude of all sampling predator/parasite prokaryotes are present in the water body sites. At the same time, the diversity indices (based on (Li et al. 2014). amplicon sequencing) showed the bacterial communities Actinobacteria were also detected earlier in Lake Bala- are less diverse in the Tihany region. It is also true that ton (Szabó, 1982; Farkas, 1982; Tóth, 1995), they are often the DOC values as well as TON and TOP were the less at described in freshwaters and sediments of lakes. Holm- that site: if the amount of the available substrates is lower, feldt et al. (2009) demonstrated that the prevalence of the most probably their concentration is limited, and these clades of diferent Actinobacteria has changed in relation circumstances favour the reproduction of fewer types of to total phosphorus (TOP) and Chl A, respectively. In our organisms (Howarth and Marino 2006). studies, several actinobacterial taxa were detected, among Taxon-specifc PCRs did not show pathogenic microbes them Frankiales with the abundance of the hgcI clade. among the studied ones, but the presence of ESBL-pro- Though the exact role of these bacteria in Lake Balaton ducing bacteria at Keszthely and Máriafürdő may indicate has to be defned by later studies, it is known that they can anthropogenic efect or other faecal contamination (De play a dominant role in degradation processes. Members of hgcI clade are common and abundant in a wide range

1 3 Biologia Futura (2020) 71:383–391 389 of freshwater habitats (Warnecke et al. 2004); they have bacteria that contribute signifcantly to the biogeochemical also the potential to utilize sunlight via actinorhodopsin cycles of iron and methane (Gayner 2018). which might promote anaplerotic carbon fxation (Ghy- Members of Bathyarchaeia (phylum Crenarchaeota) are lin et al. 2014). Ghai et al. (2014) studying a freshwater generalists, which appear in various environments. They are lake in Spain by metagenomic analysis showed a remark- incorporated in Feammox processes-anaerobic ammonium able potential of their genomes to transform recalcitrant oxidation coupled to Fe(III) reduction (Rios-Del Toro et al. plant detrital material (e.g. lignin-derived compounds). 2018). Some of them are involved in anaerobic methane Moreover, they detected that the abundances of Actino- oxidation. Evans et al. (2015) presumed also the syntrophy bacteria correlate inversely to those of Cyanobacteria that between Bathyarchaeota and sulphate-reducing bacteria can be responsible for prolonged damage to freshwater (SRB) towards anaerobic oxidation of methane. ecosystems. Our results connected to other phototrophic bacteria showed that their abundance is high in the water of Lake Balaton, which is not new. Kovács et al. (2012) described Conclusions for future biology the efect of irradiance on the germination of N 2-fxing, flamentous cyanobacteria in the sediment of Lake Balaton with As a conclusion, we can state that the bacterial as well as varying phosphorous supply. They focused their studies on archaeal diversity of the water of Lake Balaton is high and the most invasive Cyanobacteria of the lake (Cylindrosper- even when many taxa are uncultivable, the novel molecular mopsis raciborskii and Aphanizomenon fos-aquae). Somo- studies can reveal their hidden diversity. Prokaryotes are gyi et al. (2016) detected the dominance of picocyanobac- organized in complex communities, they can be involved in teria in the summer and dominance of picoeukaryotes in the many diferent metabolic processes, chemotrophic as well winter period. Cyanobia (Synechococcales) which are pico- as phototrophic organisms were also detected, just as bacte- sized cyanobacteria were observed also in the open water ria involving many oxidation/reduction reactions, etc. The of Lake Fertő (Somogyi et al. 2010); we showed they are microbial communities we have revealed can be responsi- dominant in the Tihany region in the summer of 2018, and ble for many steps in the biogeochemical cycles of carbon, the abundance of these bacteria is not surprising. Felföldi nitrogen, sulphur but also that of phosphorous or iron: to et al. (2011) demonstrated also the diversity and seasonal discover it in details, further investigations are necessary. dynamics of photoautotrophic picoplankton in Lake Balaton. It is true that the diversity and abundance of bacterioplank- The members of Anaerolineales (Chlorofexi) are strictly ton depend on the phytoplanktonic primary production, but anaerobic, organotrophic organisms, they are often found heterotrophic bacteria most probably are involved also in in freshwater sediments (Wise et al. 1997) and they can degradation of the residuals of diferent chemical contami- be responsible for metabolizing even different organic nations which have been detected in the lake (Simon-Delso substrates. et al. 2015; Avar et al. 2015; Maasz et al. 2019), these can Aerobic anoxic photoheterotrophic bacteria (e.g. Rhodo- be removed by the help of these complex bacterial commu- bacterales) are a diverse group of bacteria that produce bac- nities. Though the self-purifcation of the water is strong, teriochlorophyll a in the presence of oxygen (Ruiz-González humans must consider keeping the water quality safe for et al. 2020). They are facultative photoheterotrophic bacteria further use. being able to gain energy also from the utilization of light, which gives them an advantage over obligate heterotrophic Funding Open access funding provided by Eötvös Loránd University. This work was completed in the ELTE Institutional Excellence Pro- bacteria (Szabó-Tugyi et al. 2019). gram supported by the National Research, Development and Innovation Cytophaga–Flavobacterium group and Chitinophagales Ofce (NKFIH-1157-8/2019-DT). are Gram-negative, chemoheterotrophic organisms (Kirch- man, 2002), often able to degrade polymer substrates as Data accessibility Sequence reads were deposited in the NCBI SRA cellulose, and chitin may even degrade recalcitrant organic database and are accessible through the BioProject ID SAMN14732963 (Tihany), SAMN14732959 (Máriafürdő) and SAMN14732964 (Kes- substrates (Berg et al. 2009). zthely). Data of physical and chemical analysis were collected from In case of Archaea, our knowledge is still less connected the ofcial database of National Directorate of Water Management. to their possible role as most of the novelty discovered phyla do not have any cultured representatives (Castelle and Ban- Compliance with ethical standards feld 2018). On the basis of 16S rDNA sequence compari- sons, the phylum “Nanoarchaeota” represents a deep lineage Conflict of interest The authors declare that there are no conficts of within the Archaea. The predominant archaeal group of our interest connected to data of the paper. water samples belonged to Woesearchaeia; they are widely distributed in nature, anaerobic, fermentative, syntrophic

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Ethical statement Our paper does not contain any experiments con- Evans PN, Parks DH, Chadwick GL, Robbins SJ, Orphan VJ, Gold- nected to humans or animals. ing SD, Tyson GW (2015) Methane metabolism in the archaeal phylum Bathyarchaeota revealed by genome-centric metagen- omics. Science 350:434–438. https://doi.org/10.1126/scien Open Access This article is licensed under a Creative Commons Attri- ce.aac7745 bution 4.0 International License, which permits use, sharing, adapta- Farkas I (1982) Cellulózbontó aktinomicéták a Keszthelyi-öbölben tion, distribution and reproduction in any medium or format, as long és a „Zala-hatás” bakteriológiai indikációjának kérdése. MTA as you give appropriate credit to the original author(s) and the source, Biol Oszt Közl 25:191–200 provide a link to the Creative Commons licence, and indicate if changes Felföldi T, Duleba M, Somogyi B, Vajna B, Nikolausz M, Présing M, were made. The images or other third party material in this article are Márialigeti K, Vörös L (2011) Diversity and seasonal dynamics included in the article’s Creative Commons licence, unless indicated of photoautotrophic picoplankton in Lake Balaton (Hungary). otherwise in a credit line to the material. If material is not included in Aquatic Microbiol Ecol 63:273–287. https://doi.org/10.3354/ the article’s Creative Commons licence and your intended use is not ame01501 permitted by statutory regulation or exceeds the permitted use, you will Filho da Silva LVF, Tateno AF, Velloso LF, Levi JE, Fernandes need to obtain permission directly from the copyright holder. To view a S, Bento CNO, Rodriguez JC, Ramos SRTS (2004) Identifca- copy of this licence, visit http://creativeco mmons .org/licen ses/by/4.0/ . tion of Pseudomonas aeruginosa, Burkholderia cepacia Com- plex, and Stenotrophomonas maltophilia in respiratory samples from cystic fbrosis patients using multiplex PCR. 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